Purpose: to make 10 milliliters (mL) of 5 NaCl solution, to make 100mL of TE buffer: 10 mM TRIS, 1 mM EDTA (DNA storage solution), to find out if DNA an be spooled out of solution, to find out what DNA looks like, to find out its many unique properties, to find out what yield of DNA can be recovered during the isolation, to prepare and pour an agarose gel for DNA fragment analysis, to find out what the appearance of different DNA samples on an agarose gel.
Materials:
- Analytical Balance - Tabletop Milligram Balance - 7.6 x 7.6 cm Weigh Paper - 3.5 x 3.5 Weigh Boat - Lab Scoops - Sodium Chloride
-15 mL Capped Tubes - Tube Racks - TRIS - EDTA - Disodium Salt - 125 mL Bottle
- 100 mL Graduated Cylinder - pH Paper - Hydrochloric Acid - Sodium Hydroxide - Glass Rods - 50 mL Beakers
- Salmon Sperm DNA - 2 mL Pipette - P-1000 Micropipette, tips - 95% Ethanol - sharpie - 1L Tripour Plastic Peaker
- 40X TAE Buffer Concentrate - 600 mL Beakers - Agarose - 250 mL MEdia Bottle - Microwave - Hot Hands Protector
- Horizontal Gel Box - 65 degree Celcius water bath
Materials:
- Analytical Balance - Tabletop Milligram Balance - 7.6 x 7.6 cm Weigh Paper - 3.5 x 3.5 Weigh Boat - Lab Scoops - Sodium Chloride
-15 mL Capped Tubes - Tube Racks - TRIS - EDTA - Disodium Salt - 125 mL Bottle
- 100 mL Graduated Cylinder - pH Paper - Hydrochloric Acid - Sodium Hydroxide - Glass Rods - 50 mL Beakers
- Salmon Sperm DNA - 2 mL Pipette - P-1000 Micropipette, tips - 95% Ethanol - sharpie - 1L Tripour Plastic Peaker
- 40X TAE Buffer Concentrate - 600 mL Beakers - Agarose - 250 mL MEdia Bottle - Microwave - Hot Hands Protector
- Horizontal Gel Box - 65 degree Celcius water bath
procedure:
1. Make molarity calculations.
2. Create TE Solution by combining Solutions Two and Three
3. Dilute DNA with TE solution in a flask.
4. Add 2.92 grams of NaCl.
5. Add 4 mL of alcohol by trickling it down the side of the flask.
6. Spool DNA
7. Put the spooled DNA into a new tube and add 2 mL of fresh TE solution.
1. Make molarity calculations.
2. Create TE Solution by combining Solutions Two and Three
3. Dilute DNA with TE solution in a flask.
4. Add 2.92 grams of NaCl.
5. Add 4 mL of alcohol by trickling it down the side of the flask.
6. Spool DNA
7. Put the spooled DNA into a new tube and add 2 mL of fresh TE solution.
conclusion: The DNA lab ended up not working out the first time, our hypothesis of why was that the DNA diffused out and was obviously no longer traceable. We are testing this hypothesis during the week of 10/20/14. If it had worked the gel would have made it so the DNA was enclosed in something that did not react to the chemical that made the DNA inside it glow orange, unfortunately nothing turned orange in our gel.
analysis: we redid the DNA gel and it worked! You can tell that there is a lot of DNA because there is a big blob that is high up in the gel.
reflection: My group worked very well on this project, everything went smoothly aside from slight mishaps. Some of the things we could have do better is being a little more careful, though we were very careful as it is. We also had unsteady hands when using the pipepts, perhaps we should have balanced our hands on the table. Other than that the work was all distributed well and we did a good job completing the tasks.